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Updated: May 6, 2026
Author Spotlight: Advancements in Multiplex Detection of Respiratory Viruses
Published on: November 10, 2023
Simultaneous detection of influenza viruses A, B, and swine origin influenza A using multiplex one-step real-time
S H R Monavari1, H R Mollaie, M Fazlalipour
1Department of Virology and Anti Microbial Resistance Research Center, Iran University of Medical Sciences, Hemmat Highway, Tehran, Iran, hrmonavari@yahoo.com.
Abstract:
Every year, seasonal epidemics of influenza viruses are causing considerable morbidity and mortality worldwide. Also infrequent novel and rearranged strains of influenza viruses have caused quick, acute universal pandemics resulting in millions of mortalities. The usage of efficient and accurate detection is superior for infection control, effective treatment, and epidemiological supervision. Therefore, evaluation of useful real-time PCR molecular tests for the detection of pandemic viruses is important before the next wave of the pandemic. A novel quantitative real-time reverse-transcription polymerase chain reaction (qRT-PCR) assay with specific primers was used successfully for detection and monitoring of the influenza A, B, and swine influenza. The newly designed primers target highly conserved regions in influenza viruses. Our qRT-PCR assay is highly specific for detecting influenza A, B, and swine influenza viruses. The cutoff CT value was determined <38 for domestic human diagnostic test, under conditions of FDA emergency, and the reaction efficiency of the InfA, swInfA, and InfB assays were thereby estimated to be 97.9 % (R2 = 0.998), 98.3 % (R2 = 0.986), and 99.5 % (R2 = 0.995), respectively. Interestingly, based on our finding, there is no cross reactivity of detecting other viruses.
Insights
A new quantitative real-time reverse-transcription polymerase chain reaction (qRT-PCR) assay accurately detects influenza A, B, and swine influenza viruses. This molecular test is highly specific, offering efficient detection for pandemic preparedness.
Area of Science:
- Virology
- Molecular Biology
- Infectious Diseases
Background:
- Seasonal influenza epidemics and novel pandemics cause significant global morbidity and mortality.
- Accurate and efficient viral detection is crucial for infection control, treatment, and epidemiological surveillance.
- Evaluating real-time polymerase chain reaction (PCR) tests is vital for pandemic preparedness.
Purpose of the Study:
- To develop and evaluate a novel quantitative real-time reverse-transcription polymerase chain reaction (qRT-PCR) assay.
- To assess the assay's specificity and efficiency in detecting influenza A, B, and swine influenza viruses.
- To establish a reliable molecular diagnostic tool for pandemic influenza surveillance.
Main Methods:
- Designed specific primers targeting conserved regions of influenza viruses.
- Utilized a quantitative real-time reverse-transcription polymerase chain reaction (qRT-PCR) assay.
- Determined a cutoff cycle threshold (CT) value of <38 for diagnostic testing under emergency conditions.
Main Results:
- The qRT-PCR assay successfully detected and monitored influenza A, B, and swine influenza viruses.
- High reaction efficiencies were achieved: 97.9% for InfA, 98.3% for swInfA, and 99.5% for InfB.
- The assay demonstrated high specificity with no cross-reactivity observed with other viruses.
Conclusions:
- The developed qRT-PCR assay is a highly specific and efficient molecular tool for detecting influenza A, B, and swine influenza.
- This assay can aid in infection control, treatment, and epidemiological monitoring of influenza.
- The findings support the use of this qRT-PCR assay for pandemic influenza preparedness and response.
